Lukasiewicz Sylwia, Faron-Górecka Agata, Dziedzicka-Wasylewska Marta
Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
Methods Mol Biol. 2013;964:79-94. doi: 10.1007/978-1-62703-251-3_6.
The ability of certain neurotransmitter receptors to form oligomers provides an additional level of fine-tuning of intracellular signaling. Among the techniques allowing study of receptor oligomerization as well as influence of specific ligands on these processes, a biophysical approach with the use of fluorescently tagged receptors is the most sensitive. Measurement of the fluorescence resonance energy transfer (FRET) phenomenon between two fluorescently tagged receptors is considered a very useful and measurable tool to study the physical interactions between receptors either in a single cell or in a population of living cells. Here we describe the use of FRET measurement specifically to monitor protein oligomer formation between dopamine D(1)R and D(2)R, but the same methodology can be used to study other receptor proteins as well as their mutants.
某些神经递质受体形成寡聚体的能力为细胞内信号传导提供了额外的微调水平。在能够研究受体寡聚化以及特定配体对这些过程影响的技术中，使用荧光标记受体的生物物理方法最为灵敏。测量两个荧光标记受体之间的荧光共振能量转移（FRET）现象被认为是研究单个细胞或一群活细胞中受体之间物理相互作用的非常有用且可测量的工具。在此，我们描述了专门使用FRET测量来监测多巴胺D(1)R和D(2)R之间蛋白质寡聚体的形成，但相同的方法也可用于研究其他受体蛋白及其突变体。
